FVSU Research Report 2012 - Fort Valley State University

Georgia Small Ruminant Research and Extension
scientists at Fort Valley State University, in
collaboration with Drs. Steve Stice of University
of Georgia, David Donovan of U.S. Department of
Agriculture Maryland, Anil Sharma of Mayo Clinic
and Younis Abdelmoneim of Mercer University, are
developing biotechnological approaches to enhance
muscular mass (meat) to meet the future needs of
the expanding goat meat industry. Their approach
is to modify the myostatin (MSTN) gene in the
goat genome. MSTN gene product, also known as
growth differentiation factor (GDF8), is a member of
the TGF-beta family of secreted proteins. It is shown
to be involved in regulating muscle development in
most animal species.
The efficiency of transfection, as measured by
flow cytometry, was 14.5 percent after four days of
culture. The cytogenetic analysis performed on 29
G-banded metaphase cells revealed that the cell line
has a normal male goat karyotype consisting of 58
autosomes and two XY sex chromosomes, which
is consistent with earlier reports in goats. These
results suggest that GSF289 cell line with a normal
karyotype, having a high rate of proliferation and
an ability to be easily transfected with plasmid
DNA vectors, is an important tool to study genetic
manipulation of goats. These studies are continuing
to modify the myostatin gene in goat genome to
test the possibility of enhancing meat production in
goats, and possibly in other livestock.
Mutations in the MSTN gene are known to be
associated with double-muscling in cattle. Attempts
to knockout or block the expression of the MSTN
gene have resulted in enhanced muscular mass
without any noticeable adverse effects in mice.
The ultimate goal of these scientists is to test the
possibility of enhancing muscular mass (meat) in
goats by genetic manipulation of the MSTN gene.
Earlier they identified previously unknown flanking
DNA sequences of goat myostatin gene locus,
enabling them to design a gene-targeting vector to
introduce MSTN gene mutations in goat genome.
In continuation with those studies, in 2011 they
established three fibroblast cell lines (GSF289,
GSF737 and GSF2010) from ear skin explants of
normal healthy goats of Kiko and Saanen breeds.
Liquid nitrogen stocks of these frozen cells had a
viability rate of 96.2 percent in in vitro cultures.
These cells were morphologically indistinguishable
from the cell stocks prior to freezing. Analysis
of the growth of a fifth passage culture revealed
an ‘S’ shaped growth curve with a population
doubling time of 25 hours. The cell lines were found
negative for microbial, fungal and mycoplasma
contamination. The GSF289 cell line, which
originated from the Saanen breed of goats, was
further characterized for genetic transformation and
cytogenetic stability. It was successfully transfected
with pcDNA3.1/NT-GFP plasmid vector containing
the green fluorescent protein (GFP) gene under
human cytomegalovirus (CMV) promoter.
Georgia Small Ruminant Research and Extension
scientists at Fort Valley State University, in collaboration
with Drs. Steve Stice of the University of Georgia, David
Donovan of the U.S. Department of Agriculture-Maryland,
Anil Sharma of the Mayo Clinic and Younis Abdelmoneim
of Mercer University, are developing biotechnological
approaches to enhance muscular mass (meat) to meet
the future needs of the expanding goat meat industry.
Contact:
Mahipal Singh, Ph.D.
Phone: 478-825-6810
E-mail:singhm@fvsu.edu